Página 29 dos resultados de 45945 itens digitais encontrados em 0.037 segundos

‣ Protein Disulfide Isomerase Is Required for Platelet-derived Growth Factor-induced Vascular Smooth Muscle Cell Migration, Nox1 NADPH Oxidase Expression, and RhoGTPase Activation

Pescatore, Luciana A.; Bonatto, Diego; Forti, Fábio Luís; Sadok, Amine; Kovacic, Herve; Laurindo, Francisco R. M.
Fonte: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC; BETHESDA Publicador: AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC; BETHESDA
Tipo: Artigo de Revista Científica
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Vascular Smooth Muscle Cell (VSMC) migration into vessel neointima is a therapeutic target for atherosclerosis and postinjury restenosis. Nox1 NADPH oxidase-derived oxidants synergize with growth factors to support VSMC migration. We previously described the interaction between NADPH oxidases and the endoplasmic reticulum redox chaperone protein disulfide isomerase (PDI) in many cell types. However, physiological implications, as well as mechanisms of such association, are yet unclear. We show here that platelet-derived growth factor (PDGF) promoted subcellular redistribution of PDI concomitant to Nox1-dependent reactive oxygen species production and that siRNA-mediated PDI silencing inhibited such reactive oxygen species production, while nearly totally suppressing the increase in Nox1 expression, with no change in Nox4. Furthermore, PDI silencing inhibited PDGF-induced VSMC migration assessed by distinct methods, whereas PDI overexpression increased spontaneous basal VSMC migration. To address possible mechanisms of PDI effects, we searched for PDI interactome by systems biology analysis of physical protein-protein interaction networks, which indicated convergence with small GTPases and their regulator RhoGDI. PDI silencing decreased PDGF-induced Rac1 and RhoA activities...

‣ Biology of Langerhans cells: Selective migration of Langerhans cells into allogeneic and xenogeneic grafts on nude mice

Krueger, Gerald G.; Daynes, Raymond A.; Emam, Mansoor
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /03/1983 Português
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A major question challenging immunobiologists relates to those mechanisms that control the selective movement of cells involved in immune and inflammatory processes at various tissue sites such as the skin. Little is known about those influences that control the selective migration of macrophage-like Langerhans cells (LC) to normal epidermis, where it is uniformly distributed. Mechanistically, this includes the interaction of blood-borne LC precursors with the vascular endothelium of the skin and those factors that control the migration of the LC into the avascular epidermal component of the skin. By using (i) monoclonal antibodies specific for I-region associated Ia antigens found on LC from various inbred strains of animals and (ii) the congenitally athymic (nude) mouse as an immunologically compromised recipient of allografts and selected xenografts, we developed a model system to study the factors that restrict LC migration into the epidermis. Using this model, which excludes the need to lethally x-irradiate graft recipients, we established that: (i) the ingress of LC does not show major histocompatibility complex restriction [LC of the nude host are capable of migrating into the epidermis of allogeneic and certain xenogeneic (rat) skin grafts]; (ii) host LC are incapable of migrating into the epidermis of guinea pig or human skin grafts; (iii) the ingress of host LC into the epidermis of the graft is not accompanied by an overgrowth of the graft by host epidermis; and (iv) LC or LC precursors are capable of dividing in the skin or...

‣ Molecular advances in the cell biology of SARS-CoV and current disease prevention strategies

Stark, Caren J; Atreya, CD
Fonte: BioMed Central Publicador: BioMed Central
Tipo: Artigo de Revista Científica
Publicado em 15/04/2005 Português
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In the aftermath of the SARS epidemic, there has been significant progress in understanding the molecular and cell biology of SARS-CoV. Some of the milestones are the availability of viral genome sequence, identification of the viral receptor, development of an infectious cDNA clone, and the identification of viral antigens that elicit neutralizing antibodies. However, there is still a large gap in our understanding of how SARS-CoV interacts with the host cell and the rapidly changing viral genome adds another variable to this equation. Now the SARS-CoV story has entered a new phase, a search for preventive strategies and a cure for the disease. This review highlights the progress made in identifying molecular aspects of SARS-CoV biology that is relevant in developing disease prevention strategies. Authors conclude that development of successful SARS-CoV vaccines and antivirals depends on the progress we make in these areas in the immediate future.

‣ Aligning Goals, Assessments, and Activities: An Approach to Teaching PCR and Gel Electrophoresis

Phillips, Allison R.; Robertson, Amber L.; Batzli, Janet; Harris, Michelle; Miller, Sarah
Fonte: American Society for Cell Biology Publicador: American Society for Cell Biology
Tipo: Artigo de Revista Científica
Publicado em //2008 Português
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Polymerase chain reaction (PCR) and gel electrophoresis have become common techniques used in undergraduate molecular and cell biology labs. Although students enjoy learning these techniques, they often cannot fully comprehend and analyze the outcomes of their experiments because of a disconnect between concepts taught in lecture and experiments done in lab. Here we report the development and implementation of novel exercises that integrate the biological concepts of DNA structure and replication with the techniques of PCR and gel electrophoresis. Learning goals were defined based on concepts taught throughout the cell biology lab course and learning objectives specific to the PCR and gel electrophoresis lab. Exercises developed to promote critical thinking and target the underlying concepts of PCR, primer design, gel analysis, and troubleshooting were incorporated into an existing lab unit based on the detection of genetically modified organisms. Evaluative assessments for each exercise were aligned with the learning goals and used to measure student learning achievements. Our analysis found that the exercises were effective in enhancing student understanding of these concepts as shown by student performance across all learning goals. The new materials were particularly helpful in acquiring relevant knowledge...

‣ Bioengineering and Bioinformatics Summer Institutes: Meeting Modern Challenges in Undergraduate Summer Research

Butler, Peter J.; Dong, Cheng; Snyder, Alan J.; Jones, A. Daniel; Sheets, Erin D.
Fonte: American Society for Cell Biology Publicador: American Society for Cell Biology
Tipo: Artigo de Revista Científica
Publicado em //2008 Português
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Summer undergraduate research programs in science and engineering facilitate research progress for faculty and provide a close-ended research experience for students, which can prepare them for careers in industry, medicine, and academia. However, ensuring these outcomes is a challenge when the students arrive ill-prepared for substantive research or if projects are ill-defined or impractical for a typical 10-wk summer. We describe how the new Bioengineering and Bioinformatics Summer Institutes (BBSI), developed in response to a call for proposals by the National Institutes of Health (NIH) and the National Science Foundation (NSF), provide an impetus for the enhancement of traditional undergraduate research experiences with intense didactic training in particular skills and technologies. Such didactic components provide highly focused and qualified students for summer research with the goal of ensuring increased student satisfaction with research and mentor satisfaction with student productivity. As an example, we focus on our experiences with the Penn State Biomaterials and Bionanotechnology Summer Institute (PSU-BBSI), which trains undergraduates in core technologies in surface characterization, computational modeling, cell biology...

‣ Implementing Concept-based Learning in a Large Undergraduate Classroom

Morse, David; Jutras, France
Fonte: American Society for Cell Biology Publicador: American Society for Cell Biology
Tipo: Artigo de Revista Científica
Publicado em //2008 Português
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An experiment explicitly introducing learning strategies to a large, first-year undergraduate cell biology course was undertaken to see whether awareness and use of strategies had a measurable impact on student performance. The construction of concept maps was selected as the strategy to be introduced because of an inherent coherence with a course structured by concepts. Data were collected over three different semesters of an introductory cell biology course, all teaching similar course material with the same professor and all evaluated using similar examinations. The first group, used as a control, did not construct concept maps, the second group constructed individual concept maps, and the third group first constructed individual maps then validated their maps in small teams to provide peer feedback about the individual maps. Assessment of the experiment involved student performance on the final exam, anonymous polls of student perceptions, failure rate, and retention of information at the start of the following year. The main conclusion drawn is that concept maps without feedback have no significant effect on student performance, whereas concept maps with feedback produced a measurable increase in student problem-solving performance and a decrease in failure rates.

‣ Using Affinity Chromatography to Investigate Novel Protein–Protein Interactions in an Undergraduate Cell and Molecular Biology Lab Course

Belanger, Kenneth D.
Fonte: American Society for Cell Biology Publicador: American Society for Cell Biology
Tipo: Artigo de Revista Científica
Publicado em //2009 Português
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Inquiry-driven lab exercises require students to think carefully about a question, carry out an investigation of that question, and critically analyze the results of their investigation. Here, we describe the implementation and assessment of an inquiry-based laboratory exercise in which students obtain and analyze novel data that contribute to our understanding of macromolecular trafficking between the nucleus and cytoplasm in eukaryotic cells. Although many of the proteins involved in nucleocytoplasmic transport are known, the physical interactions between some of these polypeptides remain uncharacterized. In this cell and molecular biology lab exercise, students investigate novel protein–protein interactions between factors involved in nuclear RNA export. Using recombinant protein expression, protein extraction, affinity chromatography, SDS-polyacrylamide gel electrophoresis, and Western blotting, undergraduates in a sophomore-level lab course identified a previously unreported association between the soluble mRNA transport factor Mex67 and the C-terminal region of the yeast nuclear pore complex protein Nup1. This exercise immersed students in the process of investigative science, from proposing and performing experiments through analyzing data and reporting outcomes. On completion of this investigative lab sequence...

‣ ADF/Cofilin: A Functional Node in Cell Biology

Bernstein, Barbara W.; Bamburg, James R.
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
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Recent findings have significantly expanded our understanding of the regulation of actin depolymerizing factor (ADF)/cofilin proteins and the profound multifaceted impact that these well-established regulators of actin dynamics have on cell biology. In this review we discuss new aspects of previously documented regulation, such as phosphorylation, but also cover novel recently established modes of regulation and new functions of ADF (also known as destrin)/cofilin. We now understand that their activity responds to a vast array of inputs far greater than previously appreciated and that these proteins not only feed back to the crucially important dynamics of actin, but also to apoptosis cascades, phospholipid metabolism, and gene expression. We argue that this ability to respond to physiological changes by modulating those same changes makes the ADF/cofilin protein family a homeostatic regulator or ‘functional node’ in cell biology.

‣ Cdk1 Activity Is Required for Mitotic Activation of Aurora A during G2/M Transition of Human Cells

Van Horn, Robert D.; Chu, Shaoyou; Fan, Li; Yin, Tinggui; Du, Jian; Beckmann, Richard; Mader, Mary; Zhu, Guoxin; Toth, John; Blanchard, Kerry; Ye, Xiang S.
Fonte: American Society for Biochemistry and Molecular Biology Publicador: American Society for Biochemistry and Molecular Biology
Tipo: Artigo de Revista Científica
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In mammalian cells entry into and progression through mitosis are regulated by multiple mitotic kinases. How mitotic kinases interact with each other and coordinately regulate mitosis remains to be fully understood. Here we employed a chemical biology approach using selective small molecule kinase inhibitors to dissect the relationship between Cdk1 and Aurora A kinases during G2/M transition. We find that activation of Aurora A first occurs at centrosomes at late G2 and is required for centrosome separation independently of Cdk1 activity. Upon entry into mitosis, Aurora A then becomes fully activated downstream of Cdk1 activation. Inactivation of Aurora A or Plk1 individually during a synchronized cell cycle shows no significant effect on Cdk1 activation and entry into mitosis. However, simultaneous inactivation of both Aurora A and Plk1 markedly delays Cdk1 activation and entry into mitosis, suggesting that Aurora A and Plk1 have redundant functions in the feedback activation of Cdk1. Together, our data suggest that Cdk1, Aurora A, and Plk1 mitotic kinases participate in a feedback activation loop and that activation of Cdk1 initiates the feedback loop activity, leading to rapid and timely entry into mitosis in human cells. In addition...

‣ Calmodulin Controls Liver Proliferation via Interactions with C/EBPβ-LAP and C/EBPβ-LIP*

Orellana, Daniel; Liu, Xiaoying; Wang, Gou-Li; Jin, Jingling; Iakova, Polina; Timchenko, Nikolai A.
Fonte: American Society for Biochemistry and Molecular Biology Publicador: American Society for Biochemistry and Molecular Biology
Tipo: Artigo de Revista Científica
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A truncated isoform of C/EBPβ, C/EBPβ-LIP, is required for liver proliferation. This isoform is expressed at high levels in proliferating liver and in liver tumors. However, high levels of C/EBPβ-LIP are also observed in non-proliferating livers during acute phase response (APR). In this paper we present mechanisms by which liver regulates activities of C/EBPβ-LIP. We found that calmodulin (CaM) inhibits the ability of C/EBPβ-LIP to promote liver proliferation during APR through direct interactions. This activity of CaM is under negative control of Ca2+, which is reduced in nuclei of livers with APR, whereas it is increased in nuclei of proliferating livers. A mutant CaM, which does not interact with C/EBPβ-LIP, also fails to inhibit the growth promotion activity of C/EBPβ-LIP. Down-regulation of CaM in livers of LPS-treated mice causes liver proliferation via activation of C/EBPβ-LIP. Overexpression of C/EBPβ-LIP above levels of CaM also initiates liver proliferation in LPS-treated mice. In addition, CaM regulates transcriptional activity of another isoform of C/EBPβ, C/EBPβ-LAP, and might control liver biology through the regulation of both isoforms of C/EBPβ. In searching for molecular mechanisms by which C/EBPβ-LIP promotes cell proliferation...

‣ Angiopoietin-like Protein 3 Inhibits Lipoprotein Lipase Activity through Enhancing Its Cleavage by Proprotein Convertases*

Liu, Jun; Afroza, Huq; Rader, Daniel J.; Jin, Weijun
Fonte: American Society for Biochemistry and Molecular Biology Publicador: American Society for Biochemistry and Molecular Biology
Tipo: Artigo de Revista Científica
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Lipoprotein lipase (LPL)-mediated lipolysis of triglycerides is the first and rate-limiting step in chylomicron/very low density lipoprotein clearance at the luminal surface of the capillaries. Angiopoietin-like protein 3 (ANGPTL3) is shown to inhibit LPL activity and plays important roles in modulating lipoprotein metabolism in vivo. However, the mechanism by which it inhibits LPL activity remains poorly understood. Using cell-based analysis of the interaction between ANGPTL3, furin, proprotein convertase subtilisin/kexin type 5 (PCSK5), paired amino acid converting enzyme-4 (PACE4), and LPL, we demonstrated that the cleavage of LPL by proprotein convertases is an inactivation process, similar to that seen for endothelial lipase cleavage. At physiological concentrations and in the presence of cells, ANGPTL3 is a potent inhibitor of LPL. This action is due to the fact that ANGPTL3 can enhance LPL cleavage by endogenous furin and PACE4 but not by PCSK5. This effect is specific to LPL but not endothelial lipase. Both N- and C-terminal domains of LPL are required for ANGPTL3-enhanced cleavage, and the N-terminal domain of ANGPTL3 is sufficient to exert its effect on LPL cleavage. Moreover, ANGPTL3 enhances LPL cleavage in the presence of either heparan sulfate proteoglycans or glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1). By enhancing LPL cleavage...

‣ γ-Tocotrienol Induces Paraptosis-Like Cell Death in Human Colon Carcinoma SW620 Cells

Zhang, Jing-Shu; Li, Da-Ming; Ma, Yue; He, Ning; Gu, Qing; Wang, Feng-Shan; Jiang, Shu-Qing; Chen, Bing-Qing; Liu, Jia-Ren
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
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Colorectal cancer is one of the most serious illnesses among diagnosed cancer. As a new type of anti-cancer composition from tocotrienol-rich fraction of palm oil, γ-tocotrienol is widely used in anti-cancer research. The objectives of this study were to investigate the effects of γ-tocotrienol on human colon cancer SW620 and HCT-8 cells. We showed that treatment with different concentrations of γ-tocotrienol resulted in a dose dependent inhibition of cell growth. Cell death induced by γ-tocotrienol was mediated by a paraptosis-like cell death in SW620 and HCT-8 cells. Real-time RT-PCR and western blot analyses showed that γ-tocotrienol inhibited the expression level of β-catenin, cyclin D1 and c-jun. These data suggest that a paraptosis-like cell death induced by γ-tocotrienol in SW620 cells is associated with the suppression of the Wnt signaling pathway, which offers a novel tool for treating apoptosis-resistance colon cancer.

‣ Fulvestrant-Induced Cell Death and Proteasomal Degradation of Estrogen Receptor α Protein in MCF-7 Cells Require the CSK c-Src Tyrosine Kinase

Yeh, Wei-Lan; Shioda, Keiko; Coser, Kathryn R.; Rivizzigno, Danielle; McSweeney, Kristen R.; Shioda, Toshihiro
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
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Fulvestrant is a representative pure antiestrogen and a Selective Estrogen Receptor Down-regulator (SERD). In contrast to the Selective Estrogen Receptor Modulators (SERMs) such as 4-hydroxytamoxifen that bind to estrogen receptor α (ERα) as antagonists or partial agonists, fulvestrant causes proteasomal degradation of ERα protein, shutting down the estrogen signaling to induce proliferation arrest and apoptosis of estrogen-dependent breast cancer cells. We performed genome-wide RNAi knockdown screenings for protein kinases required for fulvestrant-induced apoptosis of the MCF-7 estrogen-dependent human breast caner cells and identified the c-Src tyrosine kinase (CSK), a negative regulator of the oncoprotein c-Src and related protein tyrosine kinases, as one of the necessary molecules. Whereas RNAi knockdown of CSK in MCF-7 cells by shRNA-expressing lentiviruses strongly suppressed fulvestrant-induced cell death, CSK knockdown did not affect cytocidal actions of 4-hydroxytamoxifen or paclitaxel, a chemotherapeutic agent. In the absence of CSK, fulvestrant-induced proteasomal degradation of ERα protein was suppressed in both MCF-7 and T47D estrogen-dependent breast cancer cells whereas the TP53-mutated T47D cells were resistant to the cytocidal action of fulvestrant in the presence or absence of CSK. MCF-7 cell sensitivities to fulvestrant-induced cell death or ERα protein degradation was not affected by small-molecular-weight inhibitors of the tyrosine kinase activity of c-Src...

‣ Lupus-Prone Mice Fail to Raise Antigen-Specific T Cell Responses to Intracellular Infection

Lieberman, Linda A.; Tsokos, George C.
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
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Systemic lupus erythematosus (SLE) is characterized by multiple cellular abnormalities culminating in the production of autoantibodies and immune complexes, resulting in tissue inflammation and organ damage. Besides active disease, the main cause of morbidity and mortality in SLE patients is infections, including those from opportunistic pathogens. To understand the failure of the immune system to fend off infections in systemic autoimmunity, we infected the lupus-prone murine strains B6.lpr and BXSB with the intracellular parasite Toxoplasma gondii and survival was monitored. Furthermore, mice were sacrificed days post infection and parasite burden and cellular immune responses such as cytokine production and cell activation were assessed. Mice from both strains succumbed to infection acutely and we observed greater susceptibility to infection in older mice. Increased parasite burden and a defective antigen-specific IFN-gamma response were observed in the lupus-prone mice. Furthermore, T cell:dendritic cell co-cultures established the presence of an intrinsic T cell defect responsible for the decreased antigen-specific response. An antigen-specific defect in IFN- gamma production prevents lupus-prone mice from clearing infection effectively. This study reveals the first cellular insight into the origin of increased susceptibility to infections in SLE disease and may guide therapeutic approaches.

‣ Hypertonic shock and the cell cycle: Identification of a stress-induced G2 delay in Saccharomyces cerevisiae

Alexander, Matthew Robert
Fonte: Universidade Rice Publicador: Universidade Rice
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Exposure of S. cerevisiae cells to an increase in external osmolarity induces a temporary growth arrest. Recovery from this stress is mediated by the accumulation of intracellular glycerol and the transcription of several stress response genes. This study demonstrates that an additional effect is the triggering of a cell cycle delay during G2/M. Increasing the extracellular osmolarity results in an accumulation of 1N and 2N cells, and a depletion of S phase cells from an asynchronous culture. Additionally, hypertonic stress causes a decrease in mRNA from the B-type cyclin CLB2, phosphorylation of the cyclin dependent kinase Cdc28p, and inhibition of Clb2p-Cdc28p kinase activity, while Clb2 protein levels are unaffected. The osmotic stress induced G2 delay is dependent upon the kinase Swe1p. Surprisingly, this delay is not correlated with inhibition of Clb2p-Cdc28p kinase activity. Deletion of SWE1 prevents the phosphorylation of Cdc28p in response to hypertonic shock, and removes the block to cell cycle progression. Deletion of SWE1 also causes synchronized cultures stressed in G2 to accumulate cells with mislocalized nuclei. However, deletion of SWE1 does not prevent the hypertonic stress induced inhibition of Clb2p-Cdc28p kinase activity. Conversely...

‣ Stochastic Modeling and Simulation of Biological Phenomena with Applications in Population Genetics and in Cell Populations

Li, Biao
Fonte: Universidade Rice Publicador: Universidade Rice
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Stochastic modeling and simulation play important roles in population genetics, statistical genetics, cell biology, molecular biology and evolutionary theory. This thesis explores four aspects of stochastic modeling and simulation of biological phenomena with applications. Research carried out is focused on two major themes. The first one (Chapters 2 and 3) concerns application of stochastic modeling in genetics, specifically to identify biases in analysis of genetics data. Two problems that are considered are ascertainment bias in estimation of microsatellite diversity in interspecies comparisons, and sample-selection bias in comparing different methods of rare variant analysis. The second theme (Chapters 4 and 5) concerns application of Poisson and branching process models to understand various aspects of cell proliferation, using S-phase labeling. Two model systems are: transient dynamics of proliferation of neurogenic progenitors in mouse brain with emphasis on differentiation and apoptosis, and balanced growth under different assumptions concerning DNA-replication pattern. In the first part, we investigate factors that are influencing the ascertainment bias of microsatellite allele sizes and explore the impact on estimates of mutation rates. Microsatellite loci play an important role as markers for identification...

‣ MYADM regulates Rac1 targeting to ordered membranes required for cell spreading and migration

Aranda, Juan F.; Reglero-Real, Natalia; Kremer, Leonor; Marcos-Ramiro, Beatriz; Ruiz-Sáenz, Ana; Calvo, María; Enrich, Carlos; Correas, Isabel; Millán, Jaime; Alonso, Miguel A.
Fonte: The American Society of Cell Biology Publicador: The American Society of Cell Biology
Tipo: Artigo de Revista Científica
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Copyright (2011) American Institute of Physics. This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics. The following article appeared in Molecular Biology of the Cell 22.8 (2011): 1252-1262 and may be found at http://www .molbiolcell.org/cgi/doi/10.1091/mbc.E10-11-0910; Membrane organization into condensed domains or rafts provides molecular platforms for selective recruitment of proteins. Cell migration is a general process that requires spatiotemporal targeting of Rac1 to membrane rafts. The protein machinery responsible for making rafts competent to recruit Rac1 remains elusive. Some members of the MAL family of proteins are involved in specialized processes dependent on this type of membrane. Because condensed membrane domains are a general feature of the plasma membrane of all mammalian cells, we hypothesized that MAL family members with ubiquitous expression and plasma membrane distribution could be involved in the organization of membranes for cell migration. We show that myeloid-associated differentiation marker (MYADM), a protein with unique features within the MAL family, colocalizes with Rac1 in membrane protrusions at the cell surface and distributes in condensed membranes. MYADM knockdown (KD) cells had altered membrane condensation and showed deficient incorporation of Rac1 to membrane raft fractions and...

‣ Metabolic characterization of T cell lymphoblastoid, Jurkat, in the presence of respiration inhibitors in a continuous reactor

Lee, Hei Chan
Fonte: Universidade Rice Publicador: Universidade Rice
Tipo: Thesis; Text Formato: 230 p.; application/pdf
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Commercial scale mammalian cell culture systems have many technical barriers: (i) accumulation of inhibitory waste products; (ii) sensitivity to low level of shear; and (iii) cellular oxygen demand in excess of bioreactor's capability to provide oxygen without damaging cells. Because of these problems, systematic approaches for providing sufficient oxygen transfer rates to large scale mammalian cell culture systems have yet to be adequately developed. This work, however, suggests a unique approach to this problem from a different point of view. A research program has been undertaken to determine if the metabolism of mammalian cells can be manipulated to minimize or eliminate the oxygen transfer capabilities required by the cells from a given bioreactor. The respiration inhibitor, rotenone, has been added to the cultures of a T cell fusion partner, Jurkat, at levels that completely inhibit respiration. Growth of the cells, at these concentrations of rotenone in completely defined media, did not occur unless the media was supplemented with at least 1 mM pyruvate or at least 1% fetal bovine serum. Respiration inhibited cells utilized much less glutamine, respiratory fuel, and concomitantly produces much less ammonia. They grew to one million cells per ml at near normal rate but the maximum cell density was 65% that of un-inhibited cultures. Proline and a minimum amount of glutamine (0.2 mM) were found to be essential for the growth of Jurkat cells in the presence of rotenone. Intracellular thiol content measured with flow cytometry technique was found to be lower for rotenone cultures...

‣ CD13 is a novel mediator of monocytic/endothelial cell adhesion

Mina-Osorio, Paola; Winnicka, Beata; O'Conor, Catherine; Grant, Christina L.; Vogel, Lotte K.; Rodriguez-Pinto, Daniel; Holmes, Kathryn V.; Ortega, Enrique; Shapiro, Linda H.
Fonte: The Society for Leukocyte Biology Publicador: The Society for Leukocyte Biology
Tipo: Artigo de Revista Científica
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During inflammation, cell surface adhesion molecules guide the adhesion and migration of circulating leukocytes across the endothelial cells lining the blood vessels to access the site of injury. The transmembrane molecule CD13 is expressed on monocytes and endothelial cells and has been shown to mediate homotypic cell adhesion, which may imply a role for CD13 in inflammatory monocyte trafficking. Here, we show that ligation and clustering of CD13 by mAb or viral ligands potently induce myeloid cell/endothelial adhesion in a signal transduction-dependent manner involving monocytic cytoskeletal rearrangement and filopodia formation. Treatment with soluble recombinant (r)CD13 blocks this CD13-dependent adhesion, and CD13 molecules from monocytic and endothelial cells are present in the same immunocomplex, suggesting a direct participation of CD13 in the adhesive interaction. This concept is strengthened by the fact that activated monocytic cells adhere to immobilized recombinant CD13. Furthermore, treatment with anti-CD13 antibodies in a murine model of peritonitis results in a decrease in leukocyte infiltration into the peritoneum, suggesting a potential role for CD13 in leukocyte trafficking in vivo. Therefore, this work supports a new direction for CD13 biology...

‣ Reversal of an immunity associated plant cell death program by the growth regulator auxin

Suresh Gopalan
Fonte: Nature Preceedings Publicador: Nature Preceedings
Tipo: Manuscript
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One form of plant immunity against pathogens involves a rapid host programmed cell death at the site of infection accompanied by resistance, termed the hypersensitive response (HR). Here it is shown that the HR programmed cell death program initiated by the bacterial type III secretion system dependent proteinaceous elicitor harpin from Erwinia amylovora can be reversed till very late in the process by the plant growth regulator auxin. Early inhibition or late reversal of this cell death program does not affect marker genes tightly correlated with local and systemic resistance. Cross-regulation between cell death programs and growth regulators is prevalent in different kingdoms. Thus, the concept that cell death program can be reversed till late provides a framework for further investigation of such phenomena, in addition to having utility in choosing better targets and strategies for treating mammalian and agricultural diseases.